Revista de Métodos de Diagnóstico Médico
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Evaluation for Integrity of Extracted RNA by Reference Material of RNA

Junko Takahashi, Akiko Takatsu and Hitoshi Iwahashi

Physiological information that identified from RNA expression level in tissues or organs give us benefit in clinical diagnostics. After extracting RNA from blood, tissue, or organs, diagnostics were carried out by using technique for semi-quantitative estimation of specific mRNA, such as real-time PCR or DNA microarrays. Validation of RNA quality need to quantitative estimation of gene expression at the moment of sampling, because using degraded RNA will lead to incorrect results. In this study, we conducted a series of experiment to evaluate RNA integrity using the reference materials of RNA length of 1000 base pairs (refRNA1000), and compared with RIN. There are significant correlation between RIN and the amount of refRNA1000, in varying state of degradation. There are also significant correlations between the amount of refRNA1000 and endogenous gene such as Actb and Rps18 in varying state of degradation in our study. The amount of endogenous gene reflects the extent of degradation more directly than that of RIN. To monitor the extent of degradation from the experimental starting point, we may refer the reference materials of refRNA1000. Not only for clinical diagnosis, but scientists always require the validity of experimental procedure, one of the candidate methods is using reference materials of RNA such as refRNA1000